The acid-tolerant microbiota associated with plaque from initial caries and healthy tooth surfaces.

The intent of this study was to compare the inherent acid tolerance of bacteria in samples of dental plaque from tooth sites in subjects with and without initial caries. Plaque was collected from approximal surfaces showing early enamel caries and from healthy tooth surfaces in the same subjects, as well as from enamel surfaces of caries-free individuals. In addition to plating on blood agar, the plaque samples were plated directly on non-selective solid agar medium buffered to pH 7.0, 6.0, 5.5, 5.0, 4.5 and 4.0 to avoid any loss of adaptation to acid during primary isolation of plaque bacteria. The results showed that approximately 50% of the total cultivable plaque microbiota from caries, as well as healthy tooth sites, was able to grow at pH 5.5 and 1% at pH 5.0, pH values regarded as critical for the demineralization of tooth enamel. At pH 5.0, members of the genus Streptococcus were the dominant group, but mutans streptococci accounted for less than half of the streptococcal viable count. The other acid-tolerant streptococcal isolates included Streptococcus anginosus, Streptococcus constellatus, Streptococcus gordinii, Streptococcus intermedius, Streptococcus mitis, Streptococcus oralis, Streptococcus salivarius and SStreptococcus sanguis. Analysis of the results indicated that the mutans streptococci in dental plaque were highly variable with respect to acid tolerance, and that both caries and healthy sites harboured significant numbers of mutans streptococci that were not acid-tolerant.

Effects of mouthrinses with linseed extract Salinum without/with chlorhexidine on oral conditions in patients with Sjögren's syndrome. A double-blind crossover investigation.

OBJECTIVES: To study the effect of mouthrinses with salivary replacement substances on oral conditions in patients with primary Sjögren's syndrome. DESIGN: Cross-over, double-blind study. SETTING: Facilities at the Centre for Oral Health Sciences, Malmö University and at Malmö University Hospital, Malmö, Sweden. SUBJECTS: Twenty-two patients with Sjogren's syndrome. INTERVENTION: Linseed extract Salinum alone (Sal) or with addition of chlorhexidine (Sal/Chx) was used for mouthrinsing during 3-week periods of rinsings separated by a 3-week "wash-out" period. MEASUREMENTS: Recordings of percentages of sites with dental plaque and bleeding on probing, mirror friction test and microbiological analyses. Questionnaire on oral symptoms due to reduced salivation. RESULTS: Dental plaque and bleeding on probing were reduced after Sal and after Sal/Chx. Friction was reduced after both treatments. No significant differences for counts of studied microbial groups were seen after Sal but the total anaerobically cultured microorganisms and of mutans streptococci fell after Sal/Chx (p<0.05 and p<0.001). Symptoms of oral dryness improved following Sal and Sal/Chx (p<0.05 and p<0.001 respectively). Speaking problems and burning mouth symptoms improved after use of Sal (p<0.05). CONCLUSIONS: Positive effects on symptoms in patients with Sjögren's syndrome were seen after use of Salinum without or with chlorhexidine.

Dental plaque mass and acid production activity of the microbiota on teeth.

With the purpose of elucidating the effect of dental plaque thickness on the acid production activity in dental plaque, we studied the prevalence of acid anions in dental plaque of children harbouring different amount of plaque on their teeth. On two occasions, the occurrence of plaque on the dentition was scored (Silness-Löe's index), and plaque on available smooth surfaces was collected and the wet weight determined. On the first occasion, the amount of acid anions in resting plaque, and on the second occasion, the acid anions of sucrose-exposed plaque were analysed with isotachophoresis. Mean value of plaque wet weights per individual dentition was 11.7 +/- 9.0 mg (first occasion) and 11.1 +/- 8.9 mg (second occasion). Dominating anions in resting plaque were acetate < propionate < lactate and in sucrose-exposed plaque lactate < acetate < propionate. The microbial acid production activity (acid anion per mg wet plaque weight) decreased with increasing weights of the plaque mass. The findings illustrate the cellular glycolytic metabolism of plaque microorganisms in thin and thick plaque, being a consequence of the diverse environmental condition existing in these ecosystems.

Acid formation in sucrose-exposed dental plaque in relation to caries incidence in schoolchildren.

The aim was to investigate the composition and concentration of organic acids produced by plaque bacteria in vivo and its possible relation to caries development in schoolchildren. Sucrose-exposed pooled plaque from 25 healthy teenagers was collected and the levels of Streptococcus mutans and lactobacilli were estimated. The acid anions were analysed with isotachophoresis. The prevalence and incidence of dental caries during a 2-year period was recorded. Depending on the amount of plaque collected, the subjects were divided into a low (n = 12) and a high (n = 13) plaque weight group. The dominating anions in sucrose exposed plaque were lactate, acetate and propionate. For the entire study group, no association was found between the total acid concentration and caries development but for the participants with low plaque weight a positive relationship (P < 0.01) was disclosed. No correlation was found between the concentrations of acids and the number of lactobacilli or Streptococcus mutans. The present results did not suggest that the acidogenicity of sugar-exposed pooled plaque samples was a suitable indicator of caries activity in teenagers.

Oral mucous membrane flora in patients using saliva substitutes.

UNLABELLED: Salivary substitutes are sometimes valuable for elderly people and radiotherapy patients, and may be used indefinitely. It is possible that this change in the ecology may effect the oral flora. OBJECTIVE: To analyse the presence of micro-organisms on oral mucous membranes during use of saliva substitutes. DESIGN: Cross-over single-blind study. SETTING: Clinic for Maxillofacial Surgery, Malmö University Hospital and Department of Oral Microbiology, Malmö University. SUBJECTS: 19 patients with low salivary secretion who had been radiated for cancer in the head and neck region. INTERVENTION: Two saliva substitutes: linseed extract and a carboxymethyl cellulose preparation (Salinum and MAS-84) were used for 3 week periods. MEASUREMENTS: Microbial samples taken, processed and analysed. RESULTS: No differences were observed when comparing baseline values with the results after the saliva substitutes and no significant differences between the use of different agents. CONCLUSION: The study suggests that use of linseed extract and carboxymethyl cellulose preparation during periods of weeks does not influence flora commonly related to caries, periodontitis or infections in the oral mucous membranes.

The microbiota of periodontal pockets with different depths in therapy-resistant periodontitis.

This study presents the composition of the cultivable microbiota colonising periodontal pockets of different depths among 2 patient-groups classified as non-responsive (NR-group; 11 participants) or responsive (R-group; 10 participants) to periodontal treatment. Microbiological samples from three types of pocket (< 4 mm deep A-samples; 4-5 mm B-samples; > 5 mm C-samples) were analysed by cultural methods for putative periodontitis pathogens, microbial groups constituting > or = 5% of the total cultivable flora and opportunistic pathogens. Actinomyces naeslundii, A. israelii, Bacteroides forsythus, Fusobacterium spp, Porphyromonas gingivalis, Prevotella intermedia, Peptostreptococcus micros, anaerobic streptococci and facultative anaerobic streptococci were most prevalent. Actinobacillus actinomycetemcomitans, Staphylococcus aureus, enteric rods and yeasts were less prevalent. The periodontitis pathogens Bacteroides forsythus, Fusobacterium spp, Porphyromonas gingivalis, Prevotella intermedia and Peptostreptococcus micros constituted together (on average) < or = 23% of the viable counts in the A- and B-samples of both patient groups and in the C-samples of the R-group. In the C-samples of the NR-group their mean counts were 45%. Correlations were found between smoking habits and the five pathogens in the C-samples and in pooled pocket depth samples. The results show that groups of periodontopathogens should be considered a causal factor in therapy-resistant periodontitis. Further, smoking and deep pockets can enhance a shift in the balance of the subgingival microflora predisposing a site to disease and a susceptible host may be the pre-requisite to therapy-resistant periodontitis.

6-month use of 0.2% delmopinol hydrochloride in comparison with 0.2% chlorhexidine digluconate and placebo (II). Effect on plaque and salivary microflora.

This double-blind, randomised, 6-month clinical trial with parallel group design in 68 subjects with gingivitis was conducted to study the effects on the oral flora of delmopinol hydrochloride 2 mg/ml (0.2% w/v, Decapinol Mouthwash), when used for partly supervised mouthrinsing in comparison with placebo and chlorhexidine digluconate 2 mg/ml (0.2% w/v, Hibitane Dental, ICI Pharmaceuticals, UK). Apart from estimating the total cultivable microbial dental plaque flora and salivary flora, analyses were focused on bacterial groups associated with gingivitis/periodontitis and dental caries. Furthermore, the presence of staphylococci, gram-negative enteric bacteria and yeasts in saliva were evaluated. The minimal inhibitory concentration (MIC) was determined for isolates belonging to the predominating micro-organisms in samples of both dental plaque and saliva. In relation to the findings in the placebo group, the use of delmopinol during the rinsing period did not produce an undesirable shift in the bacterial populations considered to be related to dental caries or periodontal diseases. These groups remained virtually unchanged during the study. In relation to the observations in the placebo group, slight reductions in the total cultivable plaque and salivary flora were observed during the study and no change was found in the ratio total anaerobically/aerobically cultivable microbial flora. Furthermore, no increased growth in staphylococci, enteric bacteria or yeasts was observed in the saliva samples. The pattern of changes taking place in the composition of the plaque and salivary microbial flora in samples from the participants rinsing with chlorhexidine were in most aspects similar to that observed in the delmopinol group. In the delmopinol group, no microbiologically significant changes were observed over time in the MIC-values for the isolates, neither in the plaque nor in the saliva samples, which indicates that no adaptation to delmopinol had taken place during the rinsing period. Similar observations were made for the plaque isolates in samples from the participants in the chlorhexidine group. On the other hand, when gram-positive and catalase-negative cocci from the saliva samples of the latter group were tested against chlorhexidine, 4-6 times higher MIC-values were obtained at 3 and 6 months both when compared to baseline and in comparison with the other two rinsing groups (p<0.01 or p<0.05). Neither delmopinol nor chlorhexidine showed any residual effect on the studied microbial groups in the plaque and the saliva samples 3 months after the end of treatment. In conclusion, delmopinol was accompanied by a composition of the plaque and salivary flora associated with healthy conditions in the oral cavity.

Guided tissue regeneration in the treatment of human infrabony defects. Clinical, radiographical and microbiological results: a pilot study.

The aim of the study was to evaluate the clinical, radiographical and microbiological outcome after using guided tissue regeneration (GTR) with a bioabsorbable membrane, Resolut. Subjects with bilateral infrabony defects at single rooted teeth were selected. A total of 22 teeth, 2 in each 1 of 7 patients and 4 in 2 patients, with probing pocket depth > or =5 mm, 3 months after scaling, participated. At baseline, assessments of plaque and gingival indices, bleeding on probing, probing pocket depth and probing attachment level were recorded and reproducible radiographs for computer-based bone level measurements were taken. Bacterial samples were collected to investigate the presence of periodontitis-associated bacteria, e.g., Porphyromonas/Prevotella- and Fusobactrium-like micro-organisms. One tooth was randomly treated with GTR and the contralateral with an open debridement procedure as a control. Clinical, radiographical and microbiological examinations were repeated 6 and 12 months postoperatively. Both procedures demonstrated a statistically significant improvement of gingival conditions, reduction of pocket depths and gain of attachment. When evaluating the differences between test and control teeth, none of the clinical parameters yielded statistical difference. Computer-based bone-level measurements showed only small differences in the majority of both test and control sites. The differences were not significant. Periodontitis-associated bacteria were present at baseline, but the appearance was not related to any specific site or patient and did not demonstrate any unwanted change in the 6- and 12-month samples. The findings suggest that the clinical, radiographical and microbiological improvements were not significantly enhanced with the GTR therapy.

Presence of cortisol in gingival crevicular fluid. A pilot study.

Cortisol is one of the primary mediators of the stress response, in the main having immunosuppressive effects. An important component of the host response in periodontal inflammation is gingival crevicular fluid (GCF), with constituents mainly derived from serum. Cortisol, like many other steroids, is present in saliva but its occurrence in GCF does not seem to be documented. Unstimulated whole saliva was collected and GCF was sampled on filter disks. The samples were analysed by a modified RIA method for serum in such a way that small volumes and low concentrations could be measured. Our findings suggest that the total concentration of cortisol in GCF might be estimated to levels below 1/10 of that in serum. However, what appears as a distinctive feature is the considerable variation of the cortisol concentrations for individual teeth. To our knowledge, this is the first time cortisol has been measured in gingival crevicular fluid, and this opens the prospects for further in vivo research.

6-month use of 0.2% delmopinol hydrochloride in comparison with 0.2% chlorhexidine digluconate and placebo. (I). Effect on plaque formation and gingivitis.

A double-blind, randomised, 6-month clinical trial with parallel group design in 149 patients with gingivitis was conducted to study the efficacy and safety of delmopinol hydrochloride 2 mg/ml (0.2% w/v, Decapinol Mouthwash) used for partly supervised mouthrinsing in comparison with chlorhexidine digluconate 2 mg/ml (0.2% w/v, Hibitane Dental, ICI Pharmaceuticals, UK) and placebo as an addition to normal oral hygiene. Assessments of efficacy were performed using the plaque index and bleeding on probing (BOP). Delmopinol showed 22% lower plaque index scores than placebo after 3 months (p<0.01) and 13% lower scores after 6 months. The corresponding figures for chlorhexidine were 38% (p<0.001) and 38% (p<0.001) after 3 and 6 months, respectively. Bleeding on probing was reduced for delmopinol in comparison with placebo by 11% after 3 months and by 18% (p<0.05) after 6 months. For chlorhexidine the corresponding figures were 18% (p<0.01) and 22% (p<0.01) after 3 and 6 months, respectively. While chlorhexidine showed greater plaque reduction than delmopinol (p<0.01 at 6 months), no statistically significant difference was reached between these two solutions regarding BOP. Both active solutions showed an increased amount of dental calculus in comparison with placebo. A transient anaesthetic sensation in the oral mucosa and taste affection were commonly reported adverse events in both the delmopinol and the chlorhexidine groups. The number of patients withdrawn from treatment due to adverse events or lack of cooperation was 7 in the chlorhexidine group, 4 in the placebo group and 1 in the delmopinol group. The results showed that rinsing with either 0.2% delmopinol hydrochloride or 0.2% chlorhexidine digluconate twice daily for 60 secs for 6 months results in less plaque formation and gingivitis than rinsing with placebo. Mouthrinsing with the 0.2% delmopinol hydrochloride solution was well accepted in this study.

Guided tissue regeneration for the treatment of intraosseous defects using a biabsorbable membrane. A controlled clinical study.

The aim of this controlled, clinical study was to evaluate guided tissue regeneration using a bioabsorbable membrane in periodontal intraosseous defects. Forty patients, each contributing one defect > or =4 mm in depth participated. The control group (18 individuals) received conventional flap therapy, while the test group (22 individuals) was treated using the bioabsorbable membrane, Guidor. Clinical assessments were made by one examiner, blinded with respect to treatment group, at baseline, 6 and 12 months following surgery. Baseline probing pocket depths of 7.7+/-1.4 mm in the membrane group and 7.6+/-1.9 mm in the control group were measured. Twelve month results showed a significant clinical attachment level gain in both control (1.1+/-1.8 mm), and membrane group (1.3+/-2.1 mm). Probing pocket depth reduction of 2.6+/-1.9 mm and 2.7+/-1.9 mm was observed in the respective groups. Bone sounding showed a non-significant gain of 0.4+/-1.8 mm and 0.6+/-1.4 mm at membrane and control sites, respectively. Radiographic evaluation confirmed these results. There were no significant differences found between treatment groups for any of the tested variables. Smoking had a negative effect on healing in both groups. In conclusion, clinical and radiographic results indicate that guided tissue regeneration using a bioabsorbable membrane at intraosseous defects did not predictably achieve greater clinical attachment level gain nor bone gain when compared to conventional flap therapy.

Therapy-resistant periodontitis. Psychosocial characteristics.

This study investigated the perspective of a stress system disorder in the pathogenesis of therapy-resistant periodontitis. The goal was to find indications that the stress-behaviour-immune system model holds as an explanatory model for the understanding of periodontal disease. 2 patient-groups were compared: one group classified as responding well to periodontal treatment (responsive-group, R-group, n=11); the other group was classified as responding less well to treatment (non-responsive-group, NR-group, n=11). Somatic and psychological factors were described as obtained by interviews and psychological testings. These findings were related to clinical data documented during the treatment of the patients. An exact logistic multivariate regression analysis was performed on a model based on variables selected by bivariate analysis (variable versus group). The results indicated that the NR-group patients displayed indications of more psychosocial strain and a more passive-dependent personality. The R-group patients displayed a more rigid personality and possibly a less stressful psychosocial situation in the past. The report highlights the possible contribution of stress factors in the context of therapy resistant periodontal disease, and the results seem to be understandable within the context of a stress system disorder perspective.

Therapy-resistant periodontitis (II). Compliance and general and dental health experiences.

In this exploratory case-control study, somatic and psychological factors are used for exploring whether the anamnestic information gives support to the hypothesis of a relationship between therapy-resistant periodontitis and stress factors. From retrospective comparisons of pocket probing depth charts, we classified 22 patients as either non-responding or responding to periodontal treatment. The non-responding patient group, in comparison with the responding patient group, showed a profile over time of less pocket probing depth (PPD) reduction in response to periodontal treatment. Information registered in each patient's file regarding periodontal registrations and dental status was gathered, and an interview was conducted aiming at collecting information on dental experiences and attitudes. Bivariate and logistic multivariate regression analyses were performed. The NR-group patients experienced significantly more unpleasant feelings towards dental procedures and a higher tendency to experience pain in connection with dental procedures. This group was found to start smoking at an older age than the R-group. One reason for this may be that smoking debut at an older age is more related to stress conditions than debut at a younger age. The report highlights the possible contribution of stress factors in periodontal disease.

Therapy-resistant periodontitis (I). Clinical and treatment characteristics.

The aim of this study was to evaluate the clinical criteria for classification of 2 different patient groups. From retrospective comparisons of pocket probing depth (PPD) charts, we classified 22 patients as either non-responding or responding to periodontal treatment. The non-responding patient group, in comparison with the responding patient group, showed a profile over time of less PPD reduction in response to periodontal treatment. Information registered in each patient's file regarding clinical periodontal conditions, as well as treatment procedure characteristics, was gathered. Bivariate analyses were performed. 3 clinical variables (frequency of mean PPD > or = 6 mm during the treatment period; at final registration; and the mobility score at the final registration) indicated that 2 separate patient groups had been identified. Although the patient groups exhibited advanced periodontitis at the beginning of treatment, both groups at the end of treatment had lost very few (1-2) teeth, probably as a result of regular supportive treatment. The characteristics of the selected clinical criteria warrant the use of these 2 patient groups in further analysis of the relation of psychosocial stress and microbiological characteristics of therapy-resistant periodontitis.

The effect of cyclosporin-A on the oral microflora at gingival sulcus of the ferret.

The effect of cyclosporin-A (CyA) on the dentogingival flora of ferrets with healthy and experimentally induced periodontal breakdown was studied. Five animals were given 10 mg/kg/d CyA. At the start of the experiments (day 0), ligatures were placed around 4 teeth in the right upper and lower jaws; corresponding contralateral teeth on the left side served as control. On days 0 and 28 (end of the experiment), microbiological samples were collected from the gingival sulcus of the experimental and the control teeth and from closely located gingival mucosa membrane. The samples were subjected to viable counts and to darkfield microscopic analyses. On day 0, facultative anaerobic rods, mainly Pasteurella spp, Alcaligenes spp, Corynebacterium spp. and Rothia spp dominated in the viable counts. No anaerobic bacteria were detected in the viable counts. On day 28 spirochetes increased in the experimental gingival sulcus samples and anaerobic bacteria appeared in most of the samples and constituted 40-60% of the total cultivable flora; Fusobacterium necrophorum and Eubacterium spp. predominated in the samples from the experimental sites. The results of the present study were compared with those of our previous investigation of ferrets not medicated with cyclosporin but also subject to experimental ligature periodontitis. Eubacterium spp. were absent in the animals not treated with cyclosporin, while this species was frequently present in the immunosuppressed ferrets. The results indicate that the presence of the large numbers of gram negative rods and of anaerobic bacteria may have enhanced the inflammatory process and further provoked the gingival overgrowth observed.

Effect of local application of delmopinol hydrochloride on developing and early established supragingival plaque in humans.

The aim of the study was to investigate the effect of delmopinol hydrochloride on the development of dental plaque and on newly established plaque. In addition, the influence of this compound on the composition of the microbiota colonizing the gingival mucous membrane was studied. 14 healthy male volunteers took part. After a 3 week pre-experimental period of intense oral hygiene, the participants refrained from all oral hygiene for 14 days. The buccal surfaces of cuspids and bicuspids on one side of the jaws were treated with a 1% aqueous solution of delmopinol hydrochloride (applied with a paint brush) 2 x a day for 7 days, while the contralateral side received placebo solution. On day 7, the application procedures were changed in that the test compound was applied on the teeth previously treated with placebo and vice versa. Plaque development was assessed clinically and by photo-based planimetric determination. The clinical recordings revealed that 89.3% of the placebo-treated surfaces displayed visible plaque on day 7, compared to 6.0% of the delmopinol hydrochloride treated surfaces. Delmopinol hydrochloride treatment of the previously placebo-treated surfaces resulted in a decrease in the number of surfaces with visible plaque from 89.3% on day 7 to 6% on day 14. These results were confirmed by the planimetric data. No significant change in the composition of the mucosal flora was observed during the experimental period. The present results indicate that delmopinol hydrochloride markedly reduces the formation of dental plaque on a clean tooth surface exposed to conditions which favour bacterial colonization. Furthermore, the substance appears to possess plaque-dissolving properties.

Dental caries and prolonged breast-feeding in 18-month-old Swedish children.

Three thousand children aged 18 months were screened for dental caries and ongoing breast-feeding at 46 child welfare centres in different parts of Sweden. Of these, 200 children were selected for a more comprehensive examination, involving investigation of dietary, toothbrushing and sucking habits, use of fluoride, and determination of salivary levels of mutans streptococci and lactobacilli. The children were divided into four groups: group 1: children with caries not being breast-fed; group 2: children with caries being breast-fed; group 3: caries-free children being breast-fed; group 4: caries-free gender- and age-matched reference children not being breast-fed. The results showed that 63 of the children (2.1%) had caries and 61 (2.0%) were still being breast-fed. Twelve (19.7%) of the 61 children still being breast-fed had caries compared with 51 (1.7%) of the 2939 children not being breast-fed; the difference was statistically significant. Children with caries and still being breast-fed had a mean defs of 5.3, and those with caries not being breast-fed 4.9; the difference was not statistically significant. Children with caries, irrespective of whether they were being breast-fed or not, had significantly higher numbers of cariogenic food intakes per day than caries-free children. Mutans streptococci were detected in 67% of the children and lactobacilli in 13%. Children with detectable mutans streptococci and lactobacilli had significantly more caries than those without. The results indicate that Swedish children with prolonged breast-feeding have a tendency to establish unsuitable dietary habits which constitutes a risk situation for developing caries at an early age.

Comparison of the effect of the linseed extract Salinum and a methyl cellulose preparation on the symptoms of dry mouth.

The effect of a linseed extract Salinum and a sodium carboxymethyl cellulose preparation called MAS-84 was compared with regard to its effect on the symptoms of dry mouth. Twenty patients with xerostomia, who had been treated for cancer in the head and neck by radiation were recruited from the clinic for maxillofacial surgery, Malmo University Hospital. Following radiation treatment the salivation was severely reduced. The symptoms of a general feeling of a dry mouth, difficulties in chewing and swallowing, taste disturbances, problems with speech and mouth burning were registered on a subjective verbal rating scale. In addition plaque index and gingival bleeding were determined. The study design was crossover and performed single blind. The experimental period was 7 weeks. The patients were randomly divided into 2 groups. One group used Salinum and the other MAS-84 for 3 weeks. The fourth week was a wash out period and for the next three weeks the patients shifted preparation. Each of the preparations was used ad libitum. Registrations of the various parameters were undertaken on days 0, 7 and 21 of the respective period. At the initial examination all patients reported considerable disturbances from mouth-dryness. These symptoms were reduced in 15 patients during the Salinum period and in 9 during the MAS-84 period. The relief was significantly more pronounced during the use of Salinum compared to that during the use of the methyl cellulose preparation. On day 21 plaque and gingival bleeding were significantly reduced during the Salinum period but not during the MAS-84 period. The results of the present study confirm those of a previous pilot study and indicate that the linseed mucilage significantly reduced the symptoms of dry mouth. This effect increased with increasing time of saliva substitute use. The linseed mucilage Salinum appeared to be a suitable saliva replacement in mouth dry patients.

Lactoferrin interaction with Actinobacillus actinomycetemcomitans.

The interaction of lactoferrin with Actinobacillus actinomycetemcomitans was examined in a 125I-labeled protein binding assay. The binding of human and bovine lactoferrins reached maximum within 1 h. Lactoferrin binding to the bacterium was pH-dependent and reversible. Scatchard analysis indicated the existence of two different types of binding sites on the bacterium, one with a high affinity constant k alpha approximately 8.8 x 10(-7) M) and the other with a low one (k alpha approximately 1.8 x 10(-6) M). Bacteria in the exponential phase of growth showed higher binding than cells in the stationary phase. Bacteria grown in medium containing serum and/or lysed erythrocytes bound lactoferrin to a lesser extent. Heat-inactivated serum, lysed erythrocytes and other proteins such as mucin and laminin inhibited lactoferrin binding to A. actinomycetemcomitans in a competitive binding assay. Sodium dodecyl sulfate polyacrylamide-gel electrophoresis and Western blot analysis of the cell envelope as well as the outer membrane of A. actinomycetemcomitans revealed lactoferrin-reactive protein bands at 29 kDa and 16.5 kDa. The 29-kDa band displayed a heat-modifiable lactoferrin-reactive form with a molecular weight of 34 kDa. Neither proteinase K-treated cell envelope nor lipopolysaccharide of this bacterium showed reactivity with lactoferrin. These data suggests a specific interaction of lactoferrin with outer membrane proteins of A. actinomycetemcomitans.